Serum NO Levels Research Articles

Baicalin Alleviates Lipopolysaccharide-Induced Liver Inflammation in Chicken by Suppressing TLR4-Mediated NF-κB Pathway.

As a kind of potent stimulus, lipopolysaccharide (LPS) has the ability to cause cell damage by activating toll-like receptor(TLR)4, then nuclear factor kappa B (NF-κB) translocates into the nucleus and changes the expression of related inflammatory genes. Baicalin is extracted from Radix Scutellariae, which possesses anti-inflammation, antioxidant and antibacterial properties. However, the effects of it on LPS-induced liver inflammation have not been fully elucidated. This study aims to investigate the anti-inflammatory effects of Baicalin on the LPS-induced liver inflammation and its underlying molecular mechanisms in chicken. The results of histopathological changes, serum biochemical analysis, NO levels and myeloperoxidase activity showed that Baicalin pretreatment ameliorated LPS-induced liver inflammation. ELISA and qPCR assays showed that Baicalin dose-dependently suppressed the production of IL-1β, IL-6, and TNF-α. Furthermore, the mRNA expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were significantly decreased by Baicalin. TLR4 is an important sensor in LPS infection. Molecular studies showed that the expression of TLR4 was inhibited by Baicalin pretreatment. In addition, Baicalin pretreatment inhibited NF-kB signaling pathway activation. All results demonstrated the protective effects of Baicalin pretreatment against LPS-induced liver inflammation in chicken via negative regulation of inflammatory mediators through the down-regulation of TLR4 expression and the inhibition of NF-kB activation.

TLR2 signal influences the iNOS/NO responses and worm development in C57BL/6J mice infected with Clonorchis sinensis

BackgroundAlthough the responses of inducible nitric oxide synthase (iNOS) and associated cytokine after Clonorchis sinensis infection have been studied recently, their mechanisms remain incompletely understood. In this study, we investigated the effects of toll-like receptor 2 (TLR2) signals on iNOS/nitric oxide (NO) responses after C. sinensis infection. We also evaluated the correlations between iNOS responses and worm development, which are possibly regulated by TLR2 signal.MethodsTLR2 wild-type and mutant C57BL/6 J mice were infected with 60 C. sinensis metacercariae, and the samples were collected at 30, 60, 90 and 120 days post-infection (dpi). The total serum NO levels were detected using Griess reagent after nitrate was reduced to nitrite. Hepatic tissue samples from the infected mice were sliced and stained with hematoxylin and eosin (HE) to observe worm development in the intrahepatic bile ducts. The iNOS mRNA transcripts in the splenocytes were examined by real time reverse transcriptase polymerase chain reaction (qRT-PCR), and iNOS expression was detected by immunohistochemistry.ResultsDeveloping C. sinensis juvenile worms were more abundant in the intrahepatic bile ducts of TLR2 mutant mice than those of TLR2 wild-type mice. However, no eggs were found in the faeces of both mice samples. The serum levels of total NO significantly increased in TLR2 mutant mice infected with C. sinensis at 30 (t(5) = 2.595, P = 0.049), 60 (t(5) = 7.838, P = 0.001) and 90 dpi (t(5) = 3.032, P = 0.029). Meanwhile, no changes occurred in TLR2 wild-type mice compared with uninfected controls during the experiment. The iNOS expression in splenocytes showed unexpected higher background levels in TLR2 mutant mice than those in TLR2 wild-type mice. Furthermore, the iNOS mRNA transcripts in splenocytes were significantly increased in the TLR2 wild-type mice infected with C. sinensis at 30 (t(5) = 5.139, P = 0.004), 60 (t(5) = 6.138, P = 0.002) and 90 dpi (t(5) = 6.332, P = 0.001). However, the rising of iNOS transcripts dropped under the uninfected control level in the TLR2 mutant mice at 120 dpi (t(5) = -9.082, P < 0.0001). Both total NO and iNOS transcripts were significantly higher in the TLR2 mutant mice than those in the TLR2 wild-type mice at 30 (t(5) = 3.091/2.933, P = 0.027/0.033) and 60 dpi (t(5) = 2.667/6.331, P = 0.044/0.001), respectively. In addition, the remarkable increase of iNOS expressions was immunohistochemically detected in the splenic serial sections of TLR2 wild-type mice at 30 and 60 dpi. However, the expressions of iNOS were remarkably decreased in the splenocytes of both TLR2 wild-type and mutant mice at 120 dpi.ConclusionsThese results demonstrate that TLR2 signal plays an important role in the regulation of iNOS expression after C. sinensis infection. TLR2 signal is also beneficial to limiting worm growth and development and contributing to the susceptibility to C. sinensis in which the iNOS/NO reactions possibly participate.

Protective effects of Xinji′erkang on myocardial infarction induced cardiac injury in mice

BackgroundMyocardial infarction (MI) is a major risk factor responsible for morbidity and mortality. Xinji′erkang (XJEK) has been clinically used as an effective medication in the treatment of coronary heart disease and myocarditis. The purpose of this study was to investigate the cardioprotective effect of Xinji′erkang on MI mice.MethodsForty male mice were randomly assigned into four groups as follows (n = 10): sham, model, MI with administration of XJEK and fosinopril for four weeks. At the end of studies, hemodynamic parameters and electrocardiography (ECG) were recorded. Heart and body mass were measured and heart weight/body weight (HW/BW) ratio was calculated as index of hypertrophy. The hypertrophy of heart and aorta was examined using the hematoxylin and eosin (HE) staining, and the collagen deposition was evaluated using Van Gieson (VG) staining. Serum nitric oxide level (NO), superoxide dismutase (SOD) activity and malondialdehyde (MDA) concentration were assayed by colorimetric analysis. The expressions of endothelial NO synthetase (eNOS) expression in serum and cardiac tissues were determined using ELISA assay and immunohistochemistry. Angiotensin II (Ang II) in serum and cardiac tissues was measured using ELISA assay. Besides, tumor necrosis factor-α (TNF-α), interleukin1β (IL-1β) and interleukin10 (IL-10) were observed in cardiac tissues with ELISA assay as well.ResultsThe administration of XJEK significantly improved cardiac dysfunction and abnormal ECG with reduced HW/BW ratio and ameliorated cardiomyocyte hypertrophy and collagen deposition compared to MI, which was partly due to the decreased SOD and increased MDA in serum. Moreover, XJEK treatment also improved endothelial dysfunction (ED) with not only enhanced eNOS activities in serum and cardiac tissues and elevated NO levels in serum, but also decreased Ang II content in serum and cardiac tissues. Finally, protein expressions of pro-inflammation cytokines, TNF-α and IL-1β in the cardiac tissues with XJEK treatment were significantly decreased compared to model. On the contrary, IL-10, an anti-inflammatory cytokine concentrated in cardiac tissues was significantly enhanced compared to model.ConclusionXinji′erkang exerts cardioprotective effect on myocardial infarction in mice, which may be due to the improvement of endothelial dysfunction and the reduction of oxidative stress and inflammation response.

Clinical and videocapillaroscopic features and serum concentration of nitric oxide in patients with Raynaud’s phenomenon after Multiwave Locked System laser therapy

Purpose: The aim of the study was the assessment of the influence of MLS laser therapy on morphological changes in nailfold videocapillaroscopy (NVC), clinical features, and the serum NO level in patients with primary and secondary Raynaud’s phenomenon (RP). Materials and methods: The analysis was performed on a group of 78 patients with RP and 30 healthy volunteers, who underwent NVC examination. NO concentration was assayed using the Griess method in blood serum before and after 3-weeks of laser biostimulation. MLS wasperformed with a Laser-M6 ASA Company device,for 3 weeks with weekend breaks, using thefollowing parameters: a frequency of 1500 Hz, adose of 25 J/cm2, and a time of 2.5 minutes on onehand. Results: After 3 weeks of MLS laser therapy, thebeneficial clinical effects manifested by a decrease of duration and number of RP attacks and degree of pain score on the visual analogue scale (VAS) in patients with primary and secondary RP. Clinical improvement after MLS laser therapy was reflected in the assessment of microcirculation disorders in NVC examination. Moreover, the tendency of normalization of NO concentration in the serum of patients with primary and secondary RP may suggest a favorable effect of laser biostimulation on the regulation of processes taking part in microcirculation disorders. Conclusions: The results showed that NVC is a useful diagnostic tool in the evaluation of dynamic microvascular involvement in RP patients. MLS laser therapy has a beneficial effect in patients with primary and secondary Raynaud’s phenomenon

ATF3 protects LPS‐induced acute pancreatic inflammation via modulating NFκB‐mediated iNOS production in mice

A state of excessive endotoxin (lipopolysaccharide, LPS) in the circulation is called endotoxemia. An overproduction of proinflammatory mediators in acute pancreatic inflammation, which may lead to insulin resistance, pancreas dysfunction and even death. Activating transcription factor (ATF)‐3, a subunit belongs to ATF/cAMP responsive element‐binding protein (CREB) family of transcription factor protects against stress‐induced acute kidney injury. The previous results suggest that ATF3 contributes to the negative regulation of inflammatory cytokine gene expression by recruiting histone deacetylase (HDAC)‐1 to ATF/NFκB‐binding sites. Biological activities and pathological effects of ATF3 in sepsis is unclear. The aim of this study is to assess the possible protective effects of ATF3 in endotoxemia induced by LPS in vivo studies. ATF3‐knockout (KO) and wild‐type mice (WT) were injected with LPS (10 mg/kg, ip) to induce endotoxemia. Survival rate in mouse model of endotoxemia, circulating white blood cells (WBC) level, serum NO expression, glucose level, histology, and the mRNA expression of inflammatory factors were investigated. The decreased circulating WBC levels, increased serum NO and glucose levels were significantly obtained in KO mice after LPS induction. Furthermore, inflammatory factor expressions, macrophage infiltration and β cell damage in KO mice after LPS induction were significantly reversed by injected adeno‐associated virus (AAV) to restore ATF3 gene expression. These results demonstrated that ATF3 plays a protective role to prevent LPS‐induced pancreas inflammation in experimental sepsis.Support or Funding Informationsupported by MOST and Tzu Chi Medical Center

Antitumor effects of polysaccharide from Sargassum fusiforme against human hepatocellular carcinoma HepG2 cells

Sargassum fusiforme (Harv.) Setchel, a kind of brown algae, has been applied as a therapeutic for thousands of years. This study was designed to investigate the antitumor effects of the polysaccharide (SFPS) from S. fusiform in liver cancer. The mice inoculated with HepG2 cells were orally administrated with SFPS at the doses of 100, 200 and 400 mg/kg body weight for 28 days. The products from peritoneal macrophages and serum in HepG2-bearing mice were measured. The effect of SFPS-induced cell apoptosis was measured by flow cytometry. Meanwhile, the expression levels of Bax and Bcl-2 were detected. Furthermore, the cytotoxicity of SFPS was evaluated by CCK-8 assay. Results showed that SFPS significantly inhibited growth of human HepG2 cell-transplanted tumor in nude mice, and remarkably increased serum TNF-α, IL-1, NO and IgM levels in HepG2-bearing mice. SFPS also promoted the cytokines (IL-1 and TNF-α) secreted by peritoneal macrophages in HepG2-bearing mice. SFPS exerted a stimulatory effect on apoptosis of HepG2 cells, increased the expression of Bax, and decreased the expression of Bcl-2. The results indicated that SFPS has anti-tumor and immunomodulatory activities at the high concentration, and it could be used as a potential chemopreventative and/or adjuvant chemotherapeutic agent in liver cancer.

Rat Model of Cecal Ligation and Puncture versus Colon Ascendens Stent Peritonitis: Comparative Study for Oxidative Stress

The CLP and CASP model as important research tools have been widely utilized in septic OS. In this study, we compare the oxidative stress condition of CLP and CASP rat models. Compared with sham operation group, infection condition and oxidative stress index were significantly different in CLP and CASP groups. Compared with CLP group, there was no significant difference in natural death rate, bacterial colony positive rate, infection condition, center venous oxygen saturation, and serum levels of TNF-α in the CASP group; whereas, MDA decreased significantly at 72 hours, T-SOD increased considerably at 12 and 72 hours after operation in the CASP group. The serum NO significantly decreased at all time-points except at 12 hours post-operation. Together, our study demonstrated the similarity of oxidative stress condition between CLP and CASP models. However, CASP rats had higher serum NO level than CLP rats.

The Lymphotoxin β Receptor Is Essential for Upregulation of IFN-Induced Guanylate-Binding Proteins and Survival after Toxoplasma gondii Infection.

Lymphotoxin β receptor (LTβR) signaling plays an important role in efficient initiation of host responses to a variety of pathogens, encompassing viruses, bacteria, and protozoans via induction of the type I interferon response. The present study reveals that after Toxoplasma gondii infection, LTβR−/− mice show a substantially reduced survival rate when compared to wild-type mice. LTβR−/− mice exhibit an increased parasite load and a more pronounced organ pathology. Also, a delayed increase of serum IL-12p40 and a failure of the protective IFNγ response in LTβR−/− mice were observed. Serum NO levels in LTβR−/− animals rose later and were markedly decreased compared to wild-type animals. At the transcriptional level, LTβR−/− animals exhibited a deregulated expression profile of several cytokines known to play a role in activation of innate immunity in T. gondii infection. Importantly, expression of the IFNγ-regulated murine guanylate-binding protein (mGBP) genes was virtually absent in the lungs of LTβR−/− mice. This demonstrates clearly that the LTβR is essential for the induction of a type II IFN-mediated immune response against T. gondii. The pronounced inability to effectively upregulate host defense effector molecules such as GBPs explains the high mortality rates of LTβR−/− animals after T. gondii infection.

12 Experimental study on the effect of Perilla seed in the rat model of asthma

Objectives To investigate the effects of Perilla seed in the rat model of bronchial asthma. Methods To observe the effect of ovalbumin (OVA) on the behavior of rats, NO and IL-6 levels in serum and in the lungs were measured using aluminium hydroxide sensitisation and an OVA aerosol inhalation challenge. Histopathological changes were examined to study the effects of Perilla consumption. Results The rat model of bronchial asthma was successfully established. Compared with model group, large, medium and small doses of Perilla seed significantly increased the weight of bronchial asthma rats (p Conclusions Perilla frutescens seed has a protective effect in the rat model of bronchial asthma. Acknowledgments The study was financially supported by the Collaborative Innovation Centre for Respiratory Disease Diagnosis and Treatment and Chinese Development of Henan Province National Natural Science Foundation of China (Grant No. XTZX2015023).

Pharmacodynamic action and mechanism of Du Liang soft capsule, a traditional Chinese medicine capsule, on treating nitroglycerin-induced migraine

Ethnopharmacological relevanceDu Liang soft capsule (DL) is a traditional Chinese medicine for treating migraines; it is made from two Chinese herbs, including LigusticumstriatumDC., root; Angelica dahurica (Hoffm.) Benth. & Hook.f. ex Franch. & Sav., root. Aim of the studyIn the present study, we aimed to elucidate the pharmacodynamic action of DL and its mechanism in an animal model of migraines induced by glyceryl trinitrate (GTN). Materials and methodsSixty rats were randomly divided into six groups, including a normal control group, model control group, positive group (Sumatriptan 0.006gkg−1), and three DL groups (0.44, 1.31 and 3.93gkg−1). All rats were intragastrically treated with the corresponding treatment for 7 consecutive days, and they were subcutaneously injected with GTN (10mgkg−1) 30min after the last treatment, except in the normal control group. After model establishment, the behaviors of all rats, including head scratching, cage climbing, and the development of red ears were observed continuously by digital camera every 30min for 3h. Four hours after GTN treatment, all rats were anaesthetized and the blood and tissue samples were collected. Plasma calcitonin gene related to peptide (CGRP) and endothelin (ET) levels were measured using the radioimmunoassay method, and serum NO was determined by the colorimetric method. Afterwards, the brainstem tissues were dissected and washed with physiological saline, and divided evenly into two parts. One part was used to test the monoamine levels, including levels of 5-hydroxytryptamine (5-HT), norepinephrine (NE) and dopamine (DA), by the fluorometric method, and the other part was used to determine the nuclear factor kappaB (NF-κB) p65, nuclear c-fos, inducible nitric oxide synthase (iNOS), interleukin (IL)-1β (IL-1β), and cyclooxygenase-2 (COX-2) levels by Western blot analysis. ResultsIn the pharmacodynamic action assay, DL (1.31 and 3.93gkg−1) greatly improved the abnormal behaviors of migraine rats, including head scratching and cage climbing, and the development of red ears. In the mechanism assay, compared with the control group, the plasma CGRP and serum NO levels and the brainstem 5-HT, NE and DA levels in the DL administration groups were significantly decreased; and the plasma ET levels were remarkably increased. Moreover, down-regulation of NF-κB p65, c-fos and pro-inflammatory cytokines, including iNOS, IL-1β and COX-2 in the brainstem in the DL administration groups were observed by Western blot analysis. ConclusionsThe above results suggested that DL has a therapeutic effect on migraines, and its mechanism may be related to adjusting the level of neurotransmitters and vasoactive substances, consequently relieving neurogenic inflammation.

Elevated plasma migration inhibitory factor in hypertension–hyperlipidemia patients correlates with impaired endothelial function

Migration inhibitory factor (MIF) has been shown to be critical in the pathology of early artherosclerosis; this article aim to investigate the plasma levels of MIF in hypertension plus hyperlipidemia patients.A total of 39 hypertension plus hyperlipidemia patients without any previous treatment were enrolled (HTN-HLP). Twenty-five healthy subjects were enrolled as the healthy control group (HEALTHY). Plasma MIF was measured by ELISA; laboratory and clinical characteristics were analyzed. HUVECs were treated with pooled plasma from HTN-HLP and HEALTHY groups, and the protein levels of adhesion molecules VCAM-1 and ICAM-1 were determined by ELISA. We found that plasma MIF was significantly elevated in the HTN-HLP group. Serum NO and eNOS levels were significantly lower; serum ET-1 (endothelin) levels were significantly higher in the HTN-HLP group. Furthermore, blood pressure, baPWV (brachial–ankle pulse wave velocity), and serum ET-1 level were significantly positively; serum NO and eNOS levels were negatively correlated with plasma MIF levels. Plasma from HTN-HLP significantly stimulated VCAM-1 and ICAM-1 protein expression on the surface of HUVECs.Plasma MIF was elevated in HTN-HLP patients and correlates with impaired endothelial function.

Hemodynamic and renal effects of bosentan and losartan in 2k1c hypertensive rats.

Endothelin-1 (ET-1) and Angiotensin II (Ang II) play important roles in generatinghypertension. The present work was investigated the hemodynamic and renal effects ofboth ET-1 and Ang II on 2K1C rats. The design of this study included five groups of rats:Group 1 Sham rats, Group 2 Normotensive rats (2K1C(n)), Group 3 two kidney one cliphypertensive rats (2K1C(h)), Group 4 two kidney one clip hypertensive rats + Bosentan(2K1C(h) + Bosentan), and Group 5 two kidney one clip hypertensive rats + Losartan(2K1C(h) + Losartan). Systolic blood pressure (SBP) was significantly elevated in 2K1C(h)rats from third to sixth week after surgery compared with control rats. SBP significantlyfell in bosentan and losartan treated rats. Serum nitric oxide (NO) level in hypertensiverats was slightly decreased, whereas in losartan group it was significantly increased.Serum malonaldehyde (MDA) significantly increased in bosentan treated rats ascompared to hypertensive rats. Bosentan administration caused a markedly increased inNa+excretion rate compared with 2K1C(h) rats in week four after surgery. In conclusion,losartan is more potent than bosentan in reducing hypertension via elevation of serumNO level.

Hepatotoxicity and endothelial dysfunction induced by high choline diet and the protective effects of phloretin in mice

The involvement of choline and its metabolite trimethylamine-N-oxide (TMAO) in endothelial dysfunction and atherosclerosis has been repeatedly confirmed. Phloretin, a dihydrochalcone flavonoid usually present in apples, possesses a variety of biological activities including vascular nutrition. This study was designed to investigate whether phloretin could alleviate or prevent high choline-induced vascular endothelial dysfunction and liver injury in mice. Mice were provided with 3% high choline water and given phloretin orally daily for 10 weeks. The high choline-treated mice showed the significant dyslipidemia and hyperglycemia with the impaired liver and vascular endothelium (p < 0.01). Administration of phloretin at 200 and 400 mg/kg bw significantly reduced the choline-induced elevation of serum TC, TG, LDL-C, AST, ALT, ET-1 and TXA2 (p < 0.01), and markedly antagonized the choline-induced decrease of serum PGI2, HDL-C and NO levels. Furthermore, phloretin elevated hepatic SOD and GSH-Px activities and decreased hepatic MDA levels of the mice exposed to high choline water. Moreover, histopathological test with the H&E and Oil Red O staining of liver sections confirmed the high choline diet-caused liver steatosis and the hepatoprotective effect of phloretin. These findings suggest that high choline causes oxidative damage, and phloretin alleviate vascular endothelial dysfunction and liver injury.

English

Diacetylmorphine (heroin) has many effects on the body system; it exerts effects on cardiovascular, immune and endocrine systems. The aim of the this study is to investigate the short-term effects of low and high doses of heroin on systolic blood pressure (SBP), thyroid hormones and monocyte chemoattractant protein-1 (MCP-1). The experimental rats were divided into three groups, each with six individuals and the treatments were continued for seven days. SBP significantly reduced by heroin administration in the second dose as compared with the control group. A marked decrease in the serum NO level was also noticed after first (low) and second (high) dose of administration as compared with control group. The present results also revealed that serum MCP-1 was statistically increased in the second dose of heroin group. Statistical analysis showed that both serum T 3 and T 4 levels were reduced significantly by heroin administration. In conclusions, for the first time, our findings suggested that diacetylmorphine could affect immune system through MCP-1 elevation. As well as heroin may affect cardiac and liver functions via increasing troponin-T and bilirubin levels.

Human SR-BI and SR-BII Potentiate Lipopolysaccharide-Induced Inflammation and Acute Liver and Kidney Injury in Mice.

The class B scavenger receptors BI (SR-BI) and BII (SR-BII) are high-density lipoprotein receptors that recognize various pathogens, including bacteria and their products. It has been reported that SR-BI/II null mice are more sensitive than normal mice to endotoxin-induced inflammation and sepsis. Because the SR-BI/II knockout model demonstrates multiple immune and metabolic disorders, we investigated the role of each receptor in the LPS-induced inflammatory response and tissue damage using transgenic mice with pLiv-11-directed expression of human SR-BI (hSR-BI) or human SR-BII (hSR-BII). At 6 h after i.p. LPS injection, transgenic hSR-BI and hSR-BII mice demonstrated markedly higher serum levels of proinflammatory cytokines and 2- to 3-fold increased expression levels of inflammatory mediators in the liver and kidney, compared with wild-type (WT) mice. LPS-stimulated inducible NO synthase expression was 3- to 6-fold higher in the liver and kidney of both transgenic strains, although serum NO levels were similar in all mice. Despite the lower high-density lipoprotein plasma levels, both transgenic strains responded to LPS by a 5-fold increase of plasma corticosterone levels, which were only moderately lower than in WT animals. LPS treatment resulted in MAPK activation in tissues of all mice; however, the strongest response was detected for hepatic extracellular signal-regulated protein kinase 1 and 2 and kidney JNK of both transgenic mice. Histological examination of hepatic and renal tissue from LPS-challenged mice revealed more injury in hSR-BII, but not hSR-BI, transgenic mice versus WT controls. Our findings demonstrate that hSR-BII, and to a lesser extent hSR-BI, significantly increase LPS-induced inflammation and contribute to LPS-induced tissue injury in the liver and kidney, two major organs susceptible to LPS toxicity.

Effects of glucagon-like peptide-1 on advanced glycation endproduct-induced aortic endothelial dysfunction in streptozotocin-induced diabetic rats: possible roles of Rho kinase- and AMP kinase-mediated nuclear factor κB signaling pathways.

Interaction between advanced glycation endproducts (AGEs) and receptor for AGEs (RAGE) as well as downstream pathways leads to vascular endothelial dysfunction in diabetes. Glucagon-like peptide-1 (GLP-1) has been reported to attenuate endothelial dysfunction in the models of atherosclerosis. However, whether GLP-1 exerts protective effects on aortic endothelium in diabetic animal model and the underlying mechanisms are still not well defined. Experimental diabetes was induced through administration with combination of high-fat diet and intraperitoneal injection of streptozotocin. Rats were randomly divided into four groups, including controls, diabetes, diabetes+sitagliptin (30mg/kg/day), diabetes+exenatide (3μg/kg/12h). Eventually, endothelial damage, markers of inflammation and oxidative stress, were measured. After 12weeks administration, diabetic rats received sitagliptin and exenatide showed significant elevation of serum NO level and reduction of ET-1 as well as inflammatory cytokines levels. Moreover, sitagliptin and exenatide significantly inhibited aortic oxidative stress level and improved aortic endothelial function in diabetic rats. Importantly, these drugs inhibited the protein expression level in AGE/RAGE-induced RhoA/ROCK/NF-κB/IκBα signaling pathways and activated AMPK in diabetic aorta. Finally, the target proteins of p-eNOS, iNOS, and ET-1, which reflect endothelial function, were also changed by these drugs. Our present study indicates that sitagliptin and exenatide administrations can improve endothelial function in diabetic aorta. Of note, RAGE/RhoA/ROCK and AMPK mediated NF-κB signaling pathways may be the intervention targets of these drugs to protect aortic endothelium.

Antihypertensive and cardioprotective effects of Cerebralcare granule® on spontaneously hypertensive rats from the perspective of the gaseous triumvirate NO-CO-H2S system.

Cerebralcare granule(®) (CG) has been reported to have hypotensive effect. However, several pathways involved in the mechanism of hypotension are still unclear. This study was designed to verify the antihypertensive effect of CG and to characterize its mechanism of action, especially from the perspective of gasotrasmmiter NO/cGMP, CO/HO and H2S/CSE systems. By using the widely used in vitro model of rat isolated thoracic aortic rings, the vasorelaxant effect of CG were studied. Furthermore, we assessed the chronic hypotensive effect of CG on spontaneously hypertensive rats (SHRs) and further to explore the potential mechanisms of its antihypertensive activity. Data in the present study demonstrated that oral treatment with CG could induce a potent antihypertensive effect. CG could reduce the intima-media thickness (IMT) of thoracic aorta significantly and increase the serum NO and H2S levels. In addition, the present results indicated that CG played a critical protective role against pressure overload-induced cardiac hypertrophy. CG not only inhibited the development of cardiac hypertrophy but also improved ventricular function. In vitro, the results showed that CG induced relaxation in rat aortic rings through an endothelium-dependent pathway mediated by NO/cGMP, CO/HO and H2S/CSE systems. Taken together, the present study demonstrated that CG could induce a potent antihypertensive effect that was partly due to the improvement of endothelial function. Also CG played a critical protective role against pressure overload-induced cardiac hypertrophy. In addition, CG could induce relaxation in rat aortic rings.

Esculin improves dyslipidemia, inflammation and renal damage in streptozotocin-induced diabetic rats

BackgroundIncreasing studies have shown that dyslipidemia and inflammatory responses play important roles in the progression of microvascular diabetic complications. Esculin (ES), a coumarin derivative, was extracted from Fraxinus rhynchophylla. The present study was to evaluate the potential effects of ES on lipid metabolism, inflammation responses and renal damage in streptozotocin (STZ)-induced experimental diabetic rats and explore the possible mechanism.MethodsDiabetic rat model was established by administration high-glucose-fat diet and intraperitoneal injection of STZ 45 mg/kg. ES was administrated to diabetic rats intragastrically at 10, 30 and 90 mg/kg for 10 weeks respectively. The levels of triglycerides (TG), total cholesterol (T-CHO), low density lipoproteins (LDL), and high-density-cholesterol (HDL-C) in serum were measured. IL-1, IL-6, ICAM-1, NO, NAGL, and AGEs level in serum were detected by ELISA assay. The accumulation of AGEs in kidney tissue was examined by immunohistochemistry assay.ResultsThe results showed that ES could decrease TG, T-CHO, LDL levels in serum of diabetic rats in a dose dependent manner. ES also decreased IL-1, IL-6, ICAM-1, NO and NGAL levels in serum of diabetic rats in a dose dependent manner. Furthermore, ES at 30 and 90 mg/kg significantly decreased AGEs level in serum and alleviated AGEs accumulation in renal in diabetic rats.ConclusionsOur findings indicate that ES could improve dyslipidemia, inflammation responses, renal damage in STZ-induced diabetic rats and the possible mechanism might be associated with the inhibition of AGEs formation.

Tranilast reduces serum IL-6 and IL-13 and protects against thioacetamide-induced acute liver injury and hepatic encephalopathy

Hepatic encephalopathy is a serious neuropsychiatric disorder usually affecting either acute or chronic hepatic failure patients. Hepatic encephalopathy was replicated in a validated rat model to assess the potential protective efficacy of tranilast against experimentally induced hepatic encephalopathy. Thioacetamide injection significantly impaired hepatic synthetic, metabolic and excretory functions with significant increase in serum NO, IL-6 and IL-13 levels and negative shift in the oxidant/antioxidant balance. Most importantly, there was a significant increase in serum ammonia levels with significant astrocytes’ swelling and vacuolization; hallmarks of hepatic encephalopathy. Tranilast administration (300mg/kg, orally) for 15 days significantly improved hepatic functions, restored oxidant/antioxidant balance, reduced serum NO, IL-6 and IL-13 levels. Meanwhile, serum ammonia significantly declined with significant reduction in astrocytes’ swelling and vacuolization. Several mechanisms can be implicated in the observed hepato- and neuroprotective potentials of tranilast, such as its anti-inflammatory potential, its antioxidant potential as well as its immunomodulatory properties.

Abstract 410: Human SR-BI/SR-BII Overexpression Enhances LPS-induced Liver and Kidney Inflammation and Acute Organ Injury

SR-BI and its splice variant SR-BII are HDL receptors that recognize bacteria and other pathogens. It has been reported that the SR-BI/II knockout mouse is more sensitive to endotoxin-induced inflammation and sepsis. Since the SR-BI/II KO model demonstrates a number of immunity-related deficiencies and metabolic alterations, we investigated the role of each receptor in the LPS-induced inflammatory response and tissue damage using transgenic mice with LIV11-directed liver/kidney expression of hSR-BI or hSR-BII. 6 hours after intraperitoneal LPS injection transgenic hSR-BII and hSR-BI mice demonstrated markedly higher serum levels of pro-inflammatory cytokines/chemokines and 2- to 3-fold increased expression levels of inflammatory mediators in the liver and kidney, compared to WT mice. In line with the pro-inflammatory status of hSR-BI and hSR-BII transgenic mice, considerably (3-6-fold) higher expression of iNOS was detected in liver and kidney of these animals, while serum NO levels were similar in all groups of mice. Despite lower levels of plasma HDL, LPS-induced corticosterone was only moderately reduced (by 30%) in plasma of both groups of transgenic mice when compared to WT animals. Histological examination of tissues from LPS-challenged mice revealed hepatic and renal injury in hSR-BII transgenic mice, while no statistically significant changes in damage score were observed between the hSR-BI transgenic and WT mice. Our findings demonstrate that hSR-BII, and to a lesser extent hSR-BI, could substantially increase LPS-triggered inflammation and contribute to LPS-induced tissue injury in the liver and kidney, two major organs that are susceptible to LPS toxicity during sepsis.